FIGURE 3.

Effect of individual amino acids on blastocyst development in media of different osmolality at LD (A–C) and HD (D–F). Zygotes (20–22 h post-hCG) were cultured for 5 days in the absence of amino acids (no a.a.) or in the presence of 1 mM L-Gln, 400 μM L-Pro, 400 μM D-Pro, 400 μM Gly, 400 μM L-Ala or 1 mM Betaine (Bet) in medium with an osmolality of (A,D) 270 mOsm/kg, (B,E) 300 mOsm/kg and (C,F) 330 mOsm/kg. Embryos were analyzed from at least three independent biological replicates, each with at least 15 embryos per treatment. *P < 0.05; **P < 0.01; ***P < 0.001 indicates significant differences from the no amino acid control, analyzed by Chi-squared test.